EP1146794B1 - Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale - Google Patents

Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale Download PDF

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EP1146794B1
EP1146794B1 EP00912432A EP00912432A EP1146794B1 EP 1146794 B1 EP1146794 B1 EP 1146794B1 EP 00912432 A EP00912432 A EP 00912432A EP 00912432 A EP00912432 A EP 00912432A EP 1146794 B1 EP1146794 B1 EP 1146794B1
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Prior art keywords
tolerogenic
protein
peptides
cooh
composition
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EP1146794A1 (fr
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Rodolphe Fritsche
Sophie Pecquet
Lionel Bovetto
Françoise MAYNARD
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Societe des Produits Nestle SA
Nestle SA
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Societe des Produits Nestle SA
Nestle SA
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/01Hydrolysed proteins; Derivatives thereof
    • A61K38/012Hydrolysed proteins; Derivatives thereof from animals
    • A61K38/018Hydrolysed proteins; Derivatives thereof from animals from milk
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/341Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
    • A23J3/343Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins of dairy proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23LFOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/17Amino acids, peptides or proteins
    • A23L33/18Peptides; Protein hydrolysates
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00Drugs for immunological or allergic disorders
    • A61P37/08Antiallergic agents

Definitions

  • This invention relates to a hypoallergenic composition containing specific tolerogenic peptides of proteins, said composition can induce oral tolerance to the native proteins. It also relates to the use of tolerogenic peptides of milk protein for the preparation of a composition inducing immulogical tolerance to milk proteins.
  • Targeted food avoidance represents a hard task for adult patients with food allergy. For example, to strictly remove cow's milk from the diet of allergic infants might be even more difficult, especially if breastfeeding is not possible or desired.
  • cow's milk The allergies to cow's milk and to the formulas containing cow's milk adapted to the needs of infants are due to the fact that the proteins of cow's milk differ from the proteins of mother's milk and can constitute allergens.
  • whey proteins Among whey proteins, ⁇ -lactoglobulin is the major component and is a strong allergen.
  • hypoallergenic formulae are systematically prescribed to "at risk” newborns, namely asymptomatic infants with atopic parents.
  • cow's milk proteins have been hydrolyzed in hypoallergenic formulae, to decrease the potential allergenicity. This approach has been demonstrated to be efficient in order to prevent sensitization by native proteins present in the adapted formulae.
  • US 5,039,532 provides an improved process for the preparation of a hydrolysate of animal milk proteins substantially reduced in allergens, in which a whey product was subjected to enzymatic hydrolysis.
  • hypoallergenic formulae Two different types are proposed to high-risk babies: partially and extensively hydrolyzed formulae, differentiated by the extent of hydrolysis of the native proteins. But as is clear from the art cited above, the primary focus to date in dealing with allergies to cow's milk has been to find preparations which will not induce an allergic response, i.e. to provide non-allergenic formulations. Nonetheless, while such formulations have permitted a person allergic to cow's milk to avoid an allergic response, they do not solve the problem which is to enable a person to drink unaltered milk products.
  • EP 0 629 350 discloses the use of non-allergenic whey protein hydrolysates which are said to be capable of inducing cow's milk protein tolerance.
  • this patent application indicates that whey protein hydrolysates substantially free of allergenic proteins could be used to induce cow's milk protein tolerance in children at risk of cow's milk allergy, the present inventors found on analysing other non-allergenic whey protein hydrolysates that non-allergenicity did not necessarily translate into the ability to induce cow's milk protein tolerance. Indeed, some of the formulations exhibiting the highest degree of non-allergenicity were found to be unsuitable for inducing cow's milk protein tolerance. Altough, Lo, C.W. and Kleiman, R.E.
  • This invention provides a hypoallergenic composition for the induction of protein tolerance in at risk individuals of protein allergy containing (i) a "non-allergenic" extensively hydrolysed proteins basis and/or (ii) a free amino acid basis, said composition comprising as the active ingredient at least one tolerogenic peptide of the allergenic protein, wherein said tolerogenic peptides are present in the form of (i) isolated tolerogenic peptidic fractions of hydrolysis of proteinaceous material containing the allergenic protein and/or (ii) synthetically prepared tolerogenic peptides, in such an amount that the ratio of tolerogenic activity by residual antigenicity is at least 2 ⁇ 10 -2 .
  • This composition contains a source of nitrogen which may provide 7 to 25% of the total energy, a source of carbohydrates which may provide at least 28 to 66% of the total energy, a source of lipids which may provide at least 25 to 60% of the total energy and at least one tolerogenic peptide of the different proteins.
  • a major advantage of this composition is to induce oral tolerance in "at risk” individuals, in order to avoid eventual sensitization by use of native tolerogens.
  • the tolerogenic peptides derived from protein hydrolysis offer both hypoallergenic and tolerogenic properties and induce oral tolerance at the humoral and cellular levels.
  • This composition is particularly intended for individuals at risk of milk protein allergy.
  • Another aspect of the present invention is the use of tolerogenic peptides of milk proteins for the preparation of a hypoallergenic composition intended for mammals susceptible to cow's milk allergy.
  • tolerogenic peptides are from milk origin and particularly from ⁇ -Lactoglobulin ( ⁇ -LG), ⁇ -lactalbumin, bovin serum albumin or casein origin.
  • ⁇ -LG ⁇ -Lactoglobulin
  • ⁇ -lactalbumin ⁇ -lactalbumin
  • bovin serum albumin bovin serum albumin or casein origin.
  • tolerogenic peptides may be used in the form of peptidic fraction containing the following peptides : H 2 N-I-D-A-L-N-E-N-K-COOH, H 2 N-V-L-V-L-D-T-D-Y-K,-K-COOH or H 2 N-T-P-E-V-D-D-E-A-L-E-K-F-D-K-COOH from ⁇ -Lactoglobulin.
  • the invention provides a method for the preparation of tolerogenic peptides useful in the induction of protein tolerance in at risk individuals to protein allergy, wherein:
  • tolerance is to be understood as a state of specific immunological unresponsivness. Both humoral (antibodies) and cell-mediated (lymphocyte .7) pathways of the immune response may be suppressed by tolerance induction. A breakdown of oral tolerance is considered to be the underlying cause of food allergy.
  • allergen is to be understood as a protein or macropeptide capable of initiating allergic reactions in humans, particularly at risk infants or nurslings. Infants are considered being "at risk" of protein allergy when either one, two parents or one sibling is atopic.
  • tolerogenic peptides is to be understood as proteic fragments, corresponding to parts of the native protein, sized from 200 to 6000 Da (3 to 50 amino acids), and preferably between 500 to 3000 Da and being able to induce specific oral tolerance to native proteins.
  • non-allergenic basis is to be understood as a nitrogen source containing a well-balanced amino-acids composition.
  • non-allergenicity is defined for milk proteins as residual allergenicity of individual whey proteins not exceeding 1 ppm and as residual allergenicity of total caseins not exceeding 10 ppm.
  • the hypoallergenic composition may contain as a source of nitrogens, peptides or free amino acids and particularly from milk proteins such as whey proteins, ⁇ -lactalbumin, ⁇ -lactoglobulin, bovine serum albumin, casein acid, caseinates, or ⁇ , ⁇ , ⁇ -casein, for example.
  • the source of nitrogen can provide at least 7 to 25 % of the total energy.
  • lactose As a source of carbohydrates, lactose, saccharose, starch or maltodextrin may be used. Carbohydrates may provide at least 28 to 66 % of the total energy.
  • Vegetable oils or butter oil are preferably used as a source of lipids which may provide at least 25 to 60 % of the total energy.
  • Vitamins, oligoelements and minerals can be added in an amount sufficient to meet daily requirements.
  • composition according to the present invention comprises as the active ingredient at least one tolerogenic peptide of the allergenic protein, said tolerogenic peptide has been selected for its ability to induce oral tolerance.
  • the tolerogenic peptides can be obtained by enzymatic hydrolysis of proteinaceous material containing the allergenic proteins that are responsible for allergies in at risk individuals, followed by isolation of tolerogenic peptidic fractions. These peptidic fractions enriched in said tolerogenic peptides can be obtained by separation of the protein hydrolysate.
  • the tolerogenic peptides may also be present in the composition in the form of synthetically prepared tolerogenic peptides.
  • compositions contain an amount sufficient to induce oral tolerance which is preferably the one which allows a complete oral tolerance induction, namely the one which prevents from any reaction after DBPCFC (double blind placebo controlled food challenge) performed with cow's milk.
  • tolerogenic peptides may be present in an amount of 0.01% to 10% (nitrogen source of the protein), for example and preferably 0.1 to 0.2 % of total peptides.
  • the composition may contain tolerogenic peptides from milk origin such as ⁇ -lactoglobulin or caseins, for example.
  • Tolerogenic peptides may thus be in the form of a peptidic fraction comprising at least one of the following peptides : H 2 N-I-D-A-L-N-E-N-K-COOH, H 2 N-V-L-V-L-D-T-D-Y-K,-K-COOH or H 2 N-T-P-E-V-D-D-E-A-L-E-K-F-D-K-COOH from ⁇ -lactoglobulin.
  • a method for the preparation of tolerogenic peptides comprises the following steps:
  • the proteinaceous material to be treated may be any composition containing protein material and in particular solution or dispersion of milk proteins : whey proteins, acid whey protein, sweet whey proteins, whey protein concentrates, whey protein isolate, demineralized whey powder or caseinates, for example.
  • the protein content may vary within the range of about 70 to 95% by weight but the starting material is preferably as rich in protein as possible.
  • the proteins present in the proteinaceous material can be modified with proteolytic enzymes into protein hydrolysate having a degree of hydrolysis ( ⁇ -amino-N/Ntot) of preferably about 10-50 %.
  • the proteolytic enzymes may be for example, from animal or vegetable origins (pepsin, chymotrypsin, trypsin, intestinal mucosa extract, pancreatic extracts, chymosin, papa ⁇ n, bromelain, ficin), bacterial or fungi origins (serine and metalloproteases from Bacillus subtilis, Bacillus licheniformis, Aspergillus orysae, Aspegillus went ⁇ i and acidic proteases from Aspergillus orizae, Aspergillus went ⁇ i, Mucor miehei, Mucor pusillus, Endothia parasitica) or a combination of these.
  • concentration of proteinaceous material in solution or in suspension is preferably around 5-20% by weight and could be pasteurised before introducing proteases.
  • the ratio enzyme/protein may be 0.1-10% weight/weight and preferably of about 0.25 to 4%.
  • Hydrolysis may be conducted at a temperature of about 35°C to 65°C, during 30 minutes to 10 hours, preferably 30 min to 4 hours at pH values within the range 2.5 to 11, preferably 4.5, 7.0, 8.0, and 8.5.
  • the pH of the solution can be adjusted and regulated with citric acid, food grade HCl or NaOH, NH 4 OH, KOH, Ca (OH) 2 for instance at a concentration of 2N pure or in blend.
  • the protein hydrolysate may be submitted to a heat treatment of about 0.1 to 10 min at a temperature of about 70 to 110°C to inactivate residual enzymes (i.e. proteases).
  • the protein hydrolysate solution thus obtained can be clarified by centrifugation and/or ultrafiltration to remove insoluble and intact proteins respectively, and the clear solution is recovered. It is possible to use at industrial scale different type of membranes (spiral, tubular, flat, allow fibbers) made with different materials (minerals, polysulfone, ...) and having different cut off limits between 1.000 and 100.000 Daltons. Depending on the type of enzyme, the hydrolysis conditions and the type of membranes the modification of the tolerogenic fractions might be sufficient at this step.
  • the recovered clear hydrolysate solution can, if desired, be concentrated by evaporation to a dry solid content of 10-50% for a subsequent treatment or spray dried if enrichment in tolerogenic peptides is sufficient.
  • the protein hydrolysate solution thus obtained can be submitted to precipitation treatment by solvent, acid, or salts, for example, followed by a centrifugation.
  • concentration of hydrolysate solution increases the yield and reduces the quantities of solvent.
  • ethanol may be added to obtain a final concentration within 15-60% volume/volume at a temperature of about 4°C to 25°C.
  • a centrifugation (30 min at 4500 g) may allow to separate soluble and insoluble peptides.
  • acid phosphoric or chlorhydric, for example
  • phospho-calcic precipitation solvents can be removed by evaporation and salts by electrodialysis for instance.
  • the clear solution and the insoluble fraction are preferably recovered.
  • the protein hydrolysate solution thus obtained may be passed into a column filled with adsorption, ion exchange or hydrophobic resin at a flow rate of 0.1-4 column volumes per hour at a temperature of about 4°C to 60°C.
  • the protein hydrolysate can be concentrated to provide a solution having a dry solid content of 8-35% by weight.
  • chromatography a fraction of peptide is absorbed into the resin by passing the hydrolysate solution into a column filled with the convenient support at a rate of 0.1-4 column volumes per hour. It is possible to use at industrial scale the different types of chromatography as: ions exchange, hydrophobic interactions, reverse phases, adsorption (hydroxyapatite, active charcoal, polystyrene base hydrophobic resins%) or covalent chromatography, for example.
  • the amount of hydrolysate solution per litre of resin filled column can be as high as 5 litres with the respect to dry solids of 10%.
  • a hydrolysate solution having 20-1000g of dry solid per litre of resin is passed into the resin filled column.
  • the chromatography treatment may be carried out at a pH of about 2 to 10 preferably 6-8, for the clarified hydrolysate solution.
  • the chromatography treatment can be conducted at a temperature of about 4°C to 60°C.
  • the chromatography treatment to select tolerogenic fractions from ⁇ -lactoglobulin may consist in using:
  • the most preferred method is to treat with resin a neutral solution, in that case, no pH adjustment is required-after hydrolysis step and the salt content of the product will be lower .
  • the column can be eluted with pure water, then water containing salts, buffer, acids, bases, or organic solvents at a temperature of 4-60°C. Elution is realised step by step or by a gradient of concentration.
  • the solutions that have passed through the column are recovered. If necessary, salts, solvents, acids, bases, are removed from the recovered solution, and the recovered solutions can be concentrated to a dry solids content of 35-65% and spray dried.
  • peptides are then specific fragments corresponding to a part of the native protein sequence or to a part of the specific tryptic peptides of hydrolysed protein.
  • tolerogenic peptides can be used for the preparation of a composition inducing oral tolerance to native proteins, said composition is intended for mammals susceptible to protein allergy and particularly human and pets.
  • ⁇ -LG was digested by TPCK treated trypsin in conditions selected to provoke its mild digestion.
  • the tolerogenic properties of the resulting peptides have been assayed in an oral tolerance experimental mouse model
  • mice Female Balb/c mice were obtained from IFFA-Crédo (L'Abresle, France). They were all bred and raised on a milk free diet. The mice were 3 weeks old at the start of the experiments.
  • ⁇ -LG (220 grams) was dissolved in bi-distilled water at 5% (w/w) final concentration.
  • the ⁇ -LG was digested by TPCK treated trypsin, using an Enzyme/Substrate (E/S) ratio of 1/100(w/w) at 40°C, pH 7.6 under constant stirring. After one hour hydrolysis, same amount of enzyme was added to give a 2/100 (w/w) final E/S ratio. After 4 hours hydrolysis, the reaction was stopped by inactivation of trypsin at 85°C for 5min. Total tryptic hydrolysate of ⁇ -lactoglobulin (TTH ⁇ -LG) was then lyophilized. The digested ⁇ -LG products were separated by preparative chromatography on a cationic resin.
  • each fraction was further characterized by its peptide content using reverse phase High Performance Liquid Chromatography. Considering TTH ⁇ -LG as reference, enrichment and impoverishment of peptides in each fraction was appreciated by area detected at 214 nm using iso nitrogen injections.
  • Gavages were orally administered to the mice at age 22 days, by gastric feeding of native ⁇ -LG (5 mg/g of body weight), various amounts of TTH ⁇ -LG, or various amounts of the different ⁇ -LG peptidic fractions.
  • Control mice were fed saline water. 5 days later, all mice were immunized ⁇ -LG and OVA (Ovalbumin, gradeV, Sigma), as a non-related antigen for testing the specificity of the immune response, 21 days after systemic challenge, a Delayed Type Hypersensitivity (DTH) evaluation was done by duplicate thickness measures of the left, rear footpad prior to, and following, immunization with ⁇ -LG.
  • OVA Optayed Type Hypersensitivity
  • Serum and intestinal fluid dilutions were assayed in duplicate for anti- ⁇ -LG and anti-OVA IgE antibodies by ELISA. Pooled samples from twenty non-immunized female mice were used as negative controls in each plates. Titers were determined by calculating the dilution of the sample which gave twice the absorbance of the negative control. Titers were expressed as the log 10 of the reciprocal of the dilution.
  • Spleen cell solutions were homogenized and purified. Cells were cocultured in the presence of ⁇ -LG or of phytohaemagglutinin A. ( 3 H)Tdr (Amersham, Zurich) was added in the final 6 hours of culture and the plates were harvested and analyzed by scintillation counting. Stimulation indices were calculated as the ratio of blank-subtracted test and control values expressed as the mean cpm ( 3 H)Tdr incorporation by triplicate cultures.
  • Fractions were pooled according to their enriched peptides similarities and in proportion to their respective masses, and then tested in vivo .
  • Fig. 1B shows that specific DTH was clearly reduced in mice fed with either TTH ⁇ -LG (0.074 mm) or each of the two tolerogenic fractions (0.063 mm and 0.062 mm respectively for F2 and F(7+9)) in comparison to local hypersensitivity measured in control mice fed only with saline (0.164 mm).
  • Fig. 1C and D show that feeding mice either of the two elected ⁇ -LG fractions as well as TTH ⁇ -LG provoked a severe decrease of the proliferative response to ⁇ -LG in comparison to the one observed from splenocytes isolated from saline fed control mice. Indeed, stimulation indices were 0.225, 0.174, and 0.341, respectively for TTH ⁇ -LG, F2, and F(7+9) fed mice. As expected, the non-specific proliferative response due to PHA stimulation was not affected by the different feeding regimen.
  • Fractions F2 and the mixture F(7+9) were both tolerogenic. Antigenicity of the tolerogenic fractions F2, F7 and F9 were lower than that of the total hydrolysate. Remarkably, the antigenicity of the tolerogenic F2 was found to be 53 times lower than the antigenicity of TTH ⁇ -LG. In these two tolerogenic fractions, the sizes of the potentially tolerogenic peptides were distributed between 8 amino acids for T6, and 23 amino acids for T21, given respective molecular weights between 915 and 2719 Da.
  • the clear permeate was treated by cantionic resin conducted as followed in a final resine volume of about 180 ml.
  • the resin was washed and equilibrated with 0.1N NaOH at a flow rate of 300 ml/h. 100 ml of clear permeate 40,000 Da were passed into the column.
  • the non retained fraction was eluted by 540 ml of pure water.
  • the second and the third were eluted by 900 ml of 0.5N HCl and 540 ml of 0.1N NaOH respectively.
  • the tolerogenic peptides are present in fraction F2 for example, in an amount sufficient to prevent from any reaction after DBPCFC performed with cow's milk.
  • Example 3 obtention of tolerogenic peptides from caseinates
  • natrium caseinate 1 kg was dissolved in 7 litres of water at 50°C.
  • the pH was adjusted at 7.5 with 2N KOH.
  • the volume was adjusted to 10 litres with water at 50°C to obtain a 10% solution with respect of powder.
  • Reverse phase chromatography 300 ml of the clear permeate were passed into the column equilibrated at a flow rate of 3 ml / min. The non retained fraction was eluted with 300 ml of water, then the second fraction was eluted with 200 ml of ethanol gradient 0-20%. Finally the third fraction was eluted with 200 ml of ethanol gradient 20-40 %. The column was regenerated by 400 ml gradient ethanol 40-80 % v/v and equilibrated with 500 ml of pure water.
  • the tolerogenic peptides T6 and T17 contained in tolerogenic fraction F2 were synthetically prepared.
  • T13 contained in a non tolerogenic fraction was also synthetized as a negative control.
  • Oral tolerance induction of these petides were tested in mice as described in example 1c) except that the dose is of 20 mcg/g of body weight.
  • Results Peptides tolerized anti- ⁇ -LG IgE / control Stimulation index (rate) Stimulation index (GLM) T17 2.67/3.39 0.305 0.405 T6 3.24/3.39 0.53 0.285 T6-T17(linked) 3.40/3.39 0.92 0.81 T13 (negative control) 3.20/3.39 1.7 0.87
  • T6 or T17 induces a cellular tolerance (specific inhibition of lymphocyte proliferation). T17 also induces a humoral tolerance by significantly decreasing specific anti-b-LG IgE.
  • T13 negative control
  • T6 H 2 N-I-D-A-L-N-E-N-K-COOH
  • T17 H 2 N-V-L-V-L-D-T-D-Y-K,-K-COOH
  • T6 H 2 N-I-D-A-L-N-E-N-K-COOH
  • T17 H 2 N-V-L-V-L-D-T-D-Y-K,-K-COOH
  • composition for 100 g of powder contains 12.5 % of peptides (synthetized peptides T6 or T17 or tolerogenic peptides as prepared in example 2, represent about 0.1 to 0.2% of total peptides), 26 % of fats, 56 % carbohydrates (including lactose 39 %, maltodextrin 11 %, starch 6 %), traces of vitamins and oligoelements to meet daily requirements, 2.5 % minerals and 3 % moisture.
  • composition is an infant formula particularly intended for at risk infant to cow's milk allergy.
  • tolerogenic infant formula In order to obtain a tolerogenic infant formula, we prepare the following mixture containing for 100 ml of formula, 1.6 % peptides (tolerogenic peptides as prepared in example 2, represent about 0.1 to 0.2 % of total peptides), 3.4 % fat, 7.4 % carbohydrates (including lactose 5.2 %, maltodextrin 1.4 %, starch 0.8 %), traces of vitamins and oligoelements to meet daily requirements, 0.3 % of minerals and 79.9 % of water.
  • tolerogenic preparation of a small volume, we prepare the following mixture containing for 30ml of preparation, from about 0.003% to about 0.015% of tolerogenic peptides (tolerogenic peptides as prepared in example 2, represent 100% of total peptides), 7.4% of carbohydrates (including maltodextrin 1.4% and saccharose 6%), 92.5% of water and aroma.
  • this preparation is to be administered 1 to 5 times daily , to newborns « at risk » of cow's milk allergy either fed hypoallergenic formula or breastfed.

Abstract

L'invention concerne une composition hypoallergène permettant d'induire une tolérance protéinique chez des sujets à risque d'allergie protéinique, renfermant (i) une base protéinique 'non allergène' largement hydrolysée et/ou (ii) une base acide aminé libre. Ladite composition renferme, comme ingrédient actif, au moins une peptide tolérogénique de la protéine allergène.

Claims (7)

  1. Composition hypoallergénique pour l'induction d'une tolérance aux protéines chez des individus à risque de présenter une allergie aux protéines, contenant (i) un constituant de base formé d'une matière protéique "non allergénique" ayant subi une hydrolyse poussée et/ou (ii) un constituant de base formé d'aminoacides libres, ladite composition comprenant, comme ingrédient actif, au moins un peptide tolérogène de la protéine allergénique, lesdits peptides tolérogènes étant présents sous forme (i) de fractions peptidiques tolérogènes isolées provenant de l'hydrolyse de la matière protéique contenant la protéine allergénique et/ou (ii) de peptides tolérogènes préparés par synthèse, en une quantité telle que le rapport de l'activité tolérogène par l'antigénicité résiduelle soit d'au moins 2 x 10-2.
  2. Composition suivant la revendication 1, qui contient une quantité de peptides tolérogènes de 0,01 % à 10 %, de préférence de 0,1 % à 0,2 % de la source d'azote totale des protéines dans la composition.
  3. Composition suivant la revendication 1 ou 2, destinée à des mammifères sensibles à une allergie au lait de vache, qui contient une fraction peptidique comprenant H2N-I-D-A-L-N-E-N-K-COOH, H2N-V-L-V-L-D-T-D-Y-K,-K-COOH ou H2N-T-P-E-V-D-D-E-A-L-E-K-F-D-K-COOH provenant de la β-lactoglobuline.
  4. Composition suivant l'une quelconque des revendications 1 à 3, qui contient une source d'azote fournissant 7 à 25 % de l'énergie totale, une source de glucides fournissant 28 à 66 % de l'énergie totale, une source de lipides fournissant 25 à 60 % de l'énergie totale, des substances minérales et des vitamines pour répondre aux besoins quotidiens et au moins un desdits peptides tolérogènes qui est ajouté en une quantité efficace pour induire une tolérance orale.
  5. Peptide tolérogène H2N-I-D-A-L-N-E-N-K-COOH de la β-lactoglobuline, ayant l'aptitude à induire une tolérance orale aux protéines du lait.
  6. Peptide tolérogène H2N-V-L-V-L-D-T-D-Y-K,-K-COOH de la β-lactoglobuline, ayant l'aptitude à induire une tolérance aux protéines du lait.
  7. Peptide tolérogène H2N-T-P-E-V-D-D-E-A-L-E-K-F-D-K-COOH de la β-lactoglobuline, ayant l'aptitude à induire une tolérance orale aux protéines du lait.
EP00912432A 1999-01-19 2000-01-17 Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale Revoked EP1146794B1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP00912432A EP1146794B1 (fr) 1999-01-19 2000-01-17 Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP99200130 1999-01-19
EP99200130 1999-01-19
PCT/EP2000/000334 WO2000042863A1 (fr) 1999-01-19 2000-01-17 Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale
EP00912432A EP1146794B1 (fr) 1999-01-19 2000-01-17 Composition hypoallergene contenant des peptides tolerogeniques induisant une tolerance orale

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EP1146794A1 EP1146794A1 (fr) 2001-10-24
EP1146794B1 true EP1146794B1 (fr) 2003-08-06

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EP (1) EP1146794B1 (fr)
AR (1) AR028141A1 (fr)
AT (1) ATE246454T1 (fr)
AU (1) AU781013B2 (fr)
BR (1) BR0007597A (fr)
CA (1) CA2358423C (fr)
DE (1) DE60004325T2 (fr)
DK (1) DK1146794T3 (fr)
ES (1) ES2203435T3 (fr)
IL (1) IL144167A0 (fr)
MY (1) MY129566A (fr)
NZ (1) NZ512866A (fr)
TW (1) TWI251468B (fr)
WO (1) WO2000042863A1 (fr)
ZA (1) ZA200106787B (fr)

Families Citing this family (37)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE275831T1 (de) 1998-06-17 2004-10-15 New Zealand Dairy Board Bioaktive molke-eiweisshydrolysate
US6551636B2 (en) 1999-07-23 2003-04-22 Novozymes A/S Modification of foaming properties of proteins
WO2002016410A2 (fr) * 2000-08-21 2002-02-28 Apitope Technology (Bristol) Limited Procede de selection de peptide
NZ506866A (en) * 2000-09-11 2003-05-30 New Zealand Dairy Board Bioactive whey protein hydrolysate free of bitter flavours wherein the enzyme used is a heat labile protease
EP1379635A2 (fr) 2000-09-25 2004-01-14 Societe Des Produits Nestle S.A. Bacteries d'acide lactique pouvant reduire la tendance d'un individu a developper des reactions allergiques
DE60120903T2 (de) * 2001-03-01 2007-02-08 Société des Produits Nestlé S.A. Hypoallergene Nahrungsmittel zur Induzierung oraler Toleranz gegenüber Sojaproteinen
JP2003137804A (ja) * 2001-10-31 2003-05-14 Morinaga Milk Ind Co Ltd インターロイキン−18誘導剤
US7897186B2 (en) 2001-11-06 2011-03-01 Novozymes North America, Inc. Modified whey protein compositions having improved foaming properties
TWI317636B (en) * 2002-11-22 2009-12-01 Meiji Dairies Corp Nutritional compositions for liver disease patients or for patients underhigh levels of invasive stress
PL368179A1 (en) * 2004-05-24 2005-11-28 Sgp & Sons Ab Proteollytic method for processing proteins and protein hydrolysis products, enriched with exogenous, semi-exogenous and conditionally exogenous amino acids for making therapeutic preparation used in normal and medicinal diets
US7618669B2 (en) * 2005-06-01 2009-11-17 Mead Johnson Nutrition Company Low-lactose partially hydrolyzed infant formula
US20060286208A1 (en) * 2005-06-01 2006-12-21 Nagendra Rangavajla Methods for producing protein partial hydrolysates and infant formulas containing the same
AU2006313660B2 (en) * 2005-11-14 2012-07-19 Nestec S.A. Oral tolerance promotion with glycated proteins
EP1867237A1 (fr) * 2006-06-15 2007-12-19 Nestec S.A. Oeuf hypoallergénique
PT2031986E (pt) * 2006-06-15 2013-04-19 Nestec Sa Indução de tolerância a proteínas do ovo
DK2164349T3 (en) * 2006-08-04 2014-12-15 Shs Int Ltd protein-FORMULA
US20090297545A1 (en) * 2008-05-27 2009-12-03 Universite Laval Immunomodulatory dairy peptides and uses thereof
EP2258208A1 (fr) * 2009-06-02 2010-12-08 University of Limerick Produit de protéine avec une immunogénicité modifiée
ES2524067T3 (es) * 2009-12-04 2014-12-03 Mjn U.S. Holdings Llc Formulación nutricional que comprende un hidrolizado que contiene péptidos de leche de vaca y/o péptidos derivados del mismo para la inducción de tolerancia
WO2011150949A1 (fr) 2010-06-04 2011-12-08 N.V. Nutricia Oligosaccharides non digestibles pour l'induction d'une tolérance orale contre des protéines alimentaires
EP3097791B1 (fr) 2010-06-04 2018-05-16 N.V. Nutricia Oligosaccharides non digestibles pour l'induction de tolérance orale contre des protéines alimentaires
EP2436389A1 (fr) * 2010-10-01 2012-04-04 Nestec S.A. Hydrolysats de protéine à base de lait et formulations pour nourrissons et compositions nutritionnelles fabriquées à partir de ceux-ci
WO2012077076A1 (fr) * 2010-12-08 2012-06-14 Holt Patrick G Traitement ou prévention d'une sensibilité aux allergènes du lait
EP2489281A1 (fr) * 2011-02-17 2012-08-22 University of Limerick Hydrolysat de caséine
WO2013083140A1 (fr) 2011-12-07 2013-06-13 N.V. Nutricia Peptides de bêta-lactoglobuline pour traiter une allergie aux protéines du lait de vache
RU2528068C1 (ru) * 2013-04-04 2014-09-10 Государственное научное учреждение Всероссийский научно-исследовательский институт молочной промышленности Российской академии сельскохозяйственных наук (ГНУ ВНИМИ Россельхозакадемии) Способ получения ферментативного сывороточных белков
RU2531164C1 (ru) * 2013-06-11 2014-10-20 Государственное научное учреждение Всероссийский научно-исследовательский институт молочной промышленности Российской академии сельскохозяйственных наук (ГНУ ВНИМИ Россельхозакадемии) Способ получения низкогидролизованной пептидной композиции из белков молочной сыворотки
WO2015090347A1 (fr) * 2013-12-16 2015-06-25 Nestec S.A. Peptides nouvellement identifiés destinés à être utilisés dans l'induction d'une tolérance orale chez de jeunes mammifères
JP6851200B2 (ja) 2014-03-05 2021-03-31 ユーシービー バイオファルマ エスアールエル 多量体Fcタンパク質
WO2016148562A1 (fr) * 2015-03-18 2016-09-22 N.V. Nutricia Procédé pour induire une tolérance orale par administration d'un peptide dérivé de bêta-lactoglobuline en combinaison avec un probiotique
GB201509718D0 (en) * 2015-06-04 2015-07-22 Reacta Biotech Ltd Oral food challenge meal formulations
EP3307090B1 (fr) * 2015-06-12 2020-06-03 Société des Produits Nestlé S.A. Combinaison de peptides tolérogéniques et de tfg-b pour une utilisation dans l'induction et l'entretien de la tolérance orale chez les jeunes mammifères
US20170020950A1 (en) * 2015-07-23 2017-01-26 Mead Johnson Nutrition Company Methods for modulating kinases
US11197917B2 (en) 2017-12-01 2021-12-14 ByHeart, Inc. Formulations for nutritional support in subjects in need thereof
WO2019212330A1 (fr) 2018-04-30 2019-11-07 N.V. Nutricia Formule avec un peptide bêta-lactoglobuline spécifique
WO2019212329A1 (fr) 2018-04-30 2019-11-07 N.V. Nutricia Formule avec des peptides de bêta-lactoglobuline spécifiques
EP4054343B1 (fr) 2019-11-07 2023-12-27 N.V. Nutricia Hydrolysat étendu de protéine de lactosérum avec des peptides tolérogènes

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0321603A1 (fr) * 1987-12-23 1989-06-28 Societe Des Produits Nestle S.A. Procédé de préparation d'un hydrolysat de protéines de lactosérum et d'un aliment hypoallergéniques
GB9312369D0 (en) * 1993-06-16 1993-07-28 Sandoz Nutrition Ltd Organic compounds
JPH07138187A (ja) * 1993-11-16 1995-05-30 Nippon Ham Kk 経口免疫寛容原、経口慢性関節リウマチ治療剤及び機能性食品
ATE218812T1 (de) * 1996-09-06 2002-06-15 Nestle Sa Induktion von toleranz gegen kuhmilch

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BR0007597A (pt) 2001-10-16
ES2203435T3 (es) 2004-04-16
DE60004325D1 (de) 2003-09-11
EP1146794A1 (fr) 2001-10-24
TWI251468B (en) 2006-03-21
MY129566A (en) 2007-04-30
US6737076B2 (en) 2004-05-18
CA2358423C (fr) 2011-07-05
ZA200106787B (en) 2002-11-18
IL144167A0 (en) 2002-05-23
AR028141A1 (es) 2003-04-30
WO2000042863A1 (fr) 2000-07-27
DE60004325T2 (de) 2004-06-09
DK1146794T3 (da) 2003-10-06
AU781013B2 (en) 2005-04-28
AU3421000A (en) 2000-08-07
CA2358423A1 (fr) 2000-07-27
US20020037357A1 (en) 2002-03-28
ATE246454T1 (de) 2003-08-15
NZ512866A (en) 2002-10-25

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